Mutagenesis of plasmid DNA with hydroxylamine: isolation of mutants of multi-copy plasmids.

Molecular & general genetics : MGG ·Vol. 145 ·No. 1 ·1976-04-23 ·Pages 101-8

Humphreys GO, Willshaw GA, Smith HR, Anderson ES

Abstract

An investigation of in vitro mutagenesis of plasmid DNA with hydroxylamine is described. The treated plasmid DNA was used to transform Escherichia coli K12. Mutants of the plasmid NTP3, which codes for resistance to ampicillin and sulphonamides, were isolated and characterised. They were classified according to the reduction in level of their beta-lactamase activity. Hydroxylamine-induced mutants of NTP14 were also isolated. This plasmid codes for ampicillin resistance, synthesis of colicin E1, and the EcoRI restriction and modification enzymes. One class of mutants is lethal to the host strain at temperatures above 33 degrees C, but carrier strains grow well at 28 degrees C. There is evidence that these mutants code for a temperature-sensitive EcoRI modification activity: the lethal effect probably results from the cleavage of the host-cell DNA by the restriction enzyme at non-permissive temperatures. The possible genetic uses of the mutant plasmids for the production of hybrid plasmids in the bacterial cell are discussed.

MeSH Terms

Ampicillin/pharmacology DNA, Bacterial Escherichia coli/isolation & purification Extrachromosomal Inheritance Hydroxylamines/pharmacology Mutation Penicillin Resistance Plasmids Sulfonamides/pharmacology

Chemicals

DNA, Bacterial Hydroxylamines Sulfonamides Ampicillin

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Humphreys G O

Willshaw G A

Smith H R

Anderson E S

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27 references, click to expand

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Article Info

Journal

Molecular & general genetics : MGG

Abbr.

Mol Gen Genet

ISSN

0026-8925

DOI

10.1007/BF00331564

Published

1976-04-23

Pages

101-8

Language

English

Region

Germany

NLM ID

0125036

Subset

External Links

PubMed source DOI: 10.1007/BF00331564

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Mutagenesis of plasmid DNA with hydroxylamine: isolation of mutants of multi-copy plasmids.

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